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WO2006113590A3 - Flow-cytometric heteroduplex analysis for detection of genetic alterations - Google Patents

Flow-cytometric heteroduplex analysis for detection of genetic alterations Download PDF

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Publication number
WO2006113590A3
WO2006113590A3 PCT/US2006/014346 US2006014346W WO2006113590A3 WO 2006113590 A3 WO2006113590 A3 WO 2006113590A3 US 2006014346 W US2006014346 W US 2006014346W WO 2006113590 A3 WO2006113590 A3 WO 2006113590A3
Authority
WO
WIPO (PCT)
Prior art keywords
duplexes
genetic alterations
detection
dna
alterations
Prior art date
Application number
PCT/US2006/014346
Other languages
French (fr)
Other versions
WO2006113590B1 (en
WO2006113590A2 (en
Inventor
Gabor Szabo
Gyoergy Lustyik
Original Assignee
Cedars Sinai Medical Center
Gabor Szabo
Gyoergy Lustyik
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cedars Sinai Medical Center, Gabor Szabo, Gyoergy Lustyik filed Critical Cedars Sinai Medical Center
Priority to EP06750396A priority Critical patent/EP1869221A2/en
Priority to US11/910,040 priority patent/US20080187923A1/en
Publication of WO2006113590A2 publication Critical patent/WO2006113590A2/en
Publication of WO2006113590A3 publication Critical patent/WO2006113590A3/en
Publication of WO2006113590B1 publication Critical patent/WO2006113590B1/en

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/686—Polymerase chain reaction [PCR]

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The present invention provides methods and kits useful for the detection of genetic alterations and to diagnose a disease condition caused by a genetic alteration. The invention involves two different PCR products, one control 'unaltered' product and one sample product, which may contain 'altered' DNA, that are mixed, denatured, and allowed to re-anneal. The DNA duplexes are then treated enzymatically or chemically to cleave single stranded or mismatched DNA regions resulting from the genetic alterations. The alterations are then quantitated by measuring a decrease in a fluorescent signal resulting from removal of the fluorescent tag. Further embodiments of the invention include a second, different fluorescent tag that is used to measure non-specific degradation of the duplexes. Other embodiments of the invention include the use of avidin coated microbeads to which the duplexes may be attached for quantitation by flow cytometry. Still further embodiments include biological microbeads for this purpose.
PCT/US2006/014346 2005-04-15 2006-04-17 Flow-cytometric heteroduplex analysis for detection of genetic alterations WO2006113590A2 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP06750396A EP1869221A2 (en) 2005-04-15 2006-04-17 Flow-cytometric heteroduplex analysis for detection of genetic alterations
US11/910,040 US20080187923A1 (en) 2005-04-15 2006-04-17 Flow-Cytometric Heteroduplex Analysis for Detection of Genetic Alterations

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US67191205P 2005-04-15 2005-04-15
US60/671,912 2005-04-15

Publications (3)

Publication Number Publication Date
WO2006113590A2 WO2006113590A2 (en) 2006-10-26
WO2006113590A3 true WO2006113590A3 (en) 2007-03-22
WO2006113590B1 WO2006113590B1 (en) 2007-08-02

Family

ID=37115802

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2006/014346 WO2006113590A2 (en) 2005-04-15 2006-04-17 Flow-cytometric heteroduplex analysis for detection of genetic alterations

Country Status (3)

Country Link
US (1) US20080187923A1 (en)
EP (1) EP1869221A2 (en)
WO (1) WO2006113590A2 (en)

Families Citing this family (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3708682B1 (en) 2008-02-01 2022-11-23 The General Hospital Corporation Use of microvesicles in diagnosis and prognosis of medical diseases and conditions
WO2010070366A1 (en) 2008-12-15 2010-06-24 Debreceni Egyetem Cytometric method for the comparative analysis of the length of pcr products and uses of this method
CN102498397B (en) 2009-07-16 2016-04-06 总医院有限公司 nucleic acid analysis
US20130029339A1 (en) 2009-09-09 2013-01-31 The General Hospital Corporation Use of microvesicles in analyzing kras mutations
EP3461912B1 (en) 2009-09-09 2022-07-13 The General Hospital Corporation Use of microvesicles in analyzing nucleic acid profiles
US20140045915A1 (en) 2010-08-31 2014-02-13 The General Hospital Corporation Cancer-related biological materials in microvesicles
EP2638057B1 (en) 2010-11-10 2019-03-06 Exosome Diagnostics, Inc. Method for isolation of nucleic acid containing particles and extraction of nucleic acids therefrom
CA2835641C (en) 2011-05-11 2019-05-28 Exosome Diagnostics, Inc. Nucleic acid extraction from heterogeneous biological materials
WO2013028788A1 (en) 2011-08-22 2013-02-28 Exosome Diagnostics, Inc. Urine biomarkers
CN104995311A (en) 2012-08-30 2015-10-21 外来体诊断公司 Controls for nucleic acid assays
CA2887058C (en) 2012-10-03 2022-02-22 Exosome Diagnostics, Inc. Use of microvesicles in diagnosis, prognosis, and treatment of medical diseases and conditions
BR112016002716B1 (en) 2013-08-06 2023-02-28 Exosome Diagnostics, Inc METHOD FOR DIAGNOSING OR MONITORING PROGRESS OR RECURRENCE OF PROSTATE CANCER WITH HIGH GLEASON SCORE
JP2019513391A (en) 2016-04-15 2019-05-30 エクソサム ダイアグノスティクス，インコーポレイティド Plasma-based detection of anaplastic lymphoma kinase (ALK) nucleic acid and ALK fusion transcripts, and their use in cancer diagnosis and treatment
WO2017197399A1 (en) 2016-05-13 2017-11-16 Exosome Diagnostics, Inc. Automated and manual methods for isolation of extracellular vesicles and co-isolation of cell-free dna from biofluids
JP2019535307A (en) 2016-10-21 2019-12-12 エクソサム ダイアグノスティクス，インコーポレイティド Sequencing and analysis of exosome-bound nucleic acids
EP3548635B1 (en) 2016-11-30 2021-04-28 Exosome Diagnostics, Inc. Methods to detect mutations in plasma using exosomal rna and cell free dna from non-small cell lung cancer patients
EP3562959A2 (en) 2017-01-02 2019-11-06 Exosome Diagnostics, Inc. Methods to distinguish rna and dna in a combined preparation
CN111133106A (en) 2017-07-12 2020-05-08 外来体诊断公司 Methods for isolation and enrichment of biological fluid-derived extracellular vesicles and methods of use thereof
US12258587B2 (en) 2018-11-20 2025-03-25 Exosome Diagnostics, Inc. Compositions and methods for internal controls of microvesicle isolations

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030143605A1 (en) * 2001-12-03 2003-07-31 Si Lok Methods for the selection and cloning of nucleic acid molecules free of unwanted nucleotide sequence alterations

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20030143605A1 (en) * 2001-12-03 2003-07-31 Si Lok Methods for the selection and cloning of nucleic acid molecules free of unwanted nucleotide sequence alterations

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
"DNA Sequencing and Fragment Analysis: PASSPORT kits for EMD Assays: a novel tool for high-throughput SNP analysis and mutation scanning", GE HEALTHCARE LIFESCIENCES, April 1999 (1999-04-01), XP003009822 *
BABON J. ET AL.: "Mutation detection using fluorescent enzyme mismatch cleavage with T4 endonuclease VII", ELECTROPHORESIS, 1999, pages 1162 - 1170, XP003009821 *
BUI C.T. ET AL.: "Chemical Cleavage reactions of DNA on solid support: application in mutation detection", BMC CHEMICAL BIOLOGY, vol. 3, no. 1, May 2003 (2003-05-01), XP021017076 *
ELLIS T.P. ET AL.: "Chemical Cleavage of Mismatch: A New look at an Established Method", HUMAN MUTATION, vol. 11, 1998, pages 345 - 353, XP003009820 *
KELLAR K. ET AL.: "Multiplexed microsphere-based flow cytometric assays", EXPERIMENTAL HEMATOLOGY, vol. 30, 2002, pages 1227 - 1237, XP002326860 *

Also Published As

Publication number Publication date
EP1869221A2 (en) 2007-12-26
WO2006113590B1 (en) 2007-08-02
WO2006113590A2 (en) 2006-10-26
US20080187923A1 (en) 2008-08-07

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